Genetic validation of aminoacyl-tRNA synthetases as drug targets in Trypanosoma brucei

Savitha Kalidas, Igor Cestari, Severine Monnerat, Qiong Li, Sandesh Regmi, Nicholas Hasle, Mehdi Labaied, Marilyn Parsons, Kenneth Stuart, Margaret A. Phillips

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

Human African trypanosomiasis (HAT) is an important public health threat in sub-Saharan Africa. Current drugs are unsatisfactory, and new drugs are being sought. Few validated enzyme targets are available to support drug discovery efforts, so our goal was to obtain essentiality data on genes with proven utility as drug targets. Aminoacyl-tRNA synthetases (aaRSs) are known drug targets for bacterial and fungal pathogens and are required for protein synthesis. Here we survey the essentiality of eight Trypanosoma brucei aaRSs by RNA interference (RNAi) gene expression knockdown, covering an enzyme from each major aaRS class: valyl-tRNA synthetase (ValRS) (class Ia), tryptophanyl-tRNA synthetase (TrpRS-1) (class Ib), arginyl-tRNA synthetase (ArgRS) (class Ic), glutamyl-tRNA synthetase (GluRS) (class 1c), threonyl-tRNA synthetase (ThrRS) (class IIa), asparaginyl-tRNA synthetase (AsnRS) (class IIb), and phenylalanyl-tRNA synthetase (α and β) (PheRS) (class IIc). Knockdown of mRNA encoding these enzymes in T. brucei mammalian stage parasites showed that all were essential for parasite growth and survival in vitro. The reduced expression resulted in growth, morphological, cell cycle, and DNA content abnormalities. ThrRS was characterized in greater detail, showing that the purified recombinant enzyme displayed ThrRS activity and that the protein localized to both the cytosol and mitochondrion. Borrelidin, a known inhibitor of ThrRS, was an inhibitor of T. brucei ThrRS and showed antitrypanosomal activity. The data show that aaRSs are essential for T. brucei survival and are likely to be excellent targets for drug discovery efforts.

Original languageEnglish (US)
Pages (from-to)504-516
Number of pages13
JournalEukaryotic Cell
Volume13
Issue number4
DOIs
StatePublished - 2014

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Threonine-tRNA Ligase
Amino Acyl-tRNA Synthetases
Trypanosoma brucei brucei
asparaginyl-tRNA synthetase
Pharmaceutical Preparations
Enzymes
Drug Discovery
Arginine-tRNA Ligase
Glutamate-tRNA Ligase
Valine-tRNA Ligase
Parasites
Tryptophan-tRNA Ligase
Phenylalanine-tRNA Ligase
African Trypanosomiasis
Gene Knockdown Techniques
Africa South of the Sahara
Growth
RNA Interference
Cytosol
Cell Cycle

ASJC Scopus subject areas

  • Microbiology
  • Molecular Biology

Cite this

Genetic validation of aminoacyl-tRNA synthetases as drug targets in Trypanosoma brucei. / Kalidas, Savitha; Cestari, Igor; Monnerat, Severine; Li, Qiong; Regmi, Sandesh; Hasle, Nicholas; Labaied, Mehdi; Parsons, Marilyn; Stuart, Kenneth; Phillips, Margaret A.

In: Eukaryotic Cell, Vol. 13, No. 4, 2014, p. 504-516.

Research output: Contribution to journalArticle

Kalidas, S, Cestari, I, Monnerat, S, Li, Q, Regmi, S, Hasle, N, Labaied, M, Parsons, M, Stuart, K & Phillips, MA 2014, 'Genetic validation of aminoacyl-tRNA synthetases as drug targets in Trypanosoma brucei', Eukaryotic Cell, vol. 13, no. 4, pp. 504-516. https://doi.org/10.1128/EC.00017-14
Kalidas, Savitha ; Cestari, Igor ; Monnerat, Severine ; Li, Qiong ; Regmi, Sandesh ; Hasle, Nicholas ; Labaied, Mehdi ; Parsons, Marilyn ; Stuart, Kenneth ; Phillips, Margaret A. / Genetic validation of aminoacyl-tRNA synthetases as drug targets in Trypanosoma brucei. In: Eukaryotic Cell. 2014 ; Vol. 13, No. 4. pp. 504-516.
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